Part of the Methods in Molecular Biology book series (MIMB, volume 72)
Quantitative In Situ Hybridization Histochemistry
The key advantage of in situ hybridization histochemistry with radiolabeled as opposed to nonradiolabeled probes is that relative differences in the amount of specific mRNA transcripts present in tissue sections can be accurately and reproducibly quantified. Precise quantification and localization of transcripts, as well as the enhanced sensitivity of 35S-labeled probes, allows powerful qualitative as well as quantitative controls to be built into experiments. Interpretation of autoradiographs of tissue sections hybridized in situ depends on careful selection of control tissue and on the availability of a suitable computerized densitometer. Major limiting factors on the performance of in situ hybridization are the relative and particularly the absolute abundance of target transcripts. In situ hybridization on frozen sections produces higher signal intensity and lower background than in situ hybridization on wax-embedded sections, at the expense of tissue morphology (see Fig. 1).
KeywordsVortex Dust Microwave Chloroform Expense
© Humana Press Inc., Totowa, NJ 1997